Indirect measures of methane emissions of Sahelian zebu cattle in West Africa, role of environment and management.

In Northern Senegal, traditional cattle management systems (TRAD) which depend on natural forages coexist with intensified systems (INT) which rely on periodic supplementation with crop residues and local concentrates. This study aims to estimate the effects of seasons and management systems on the methane emissions of Gobra zebu, in relation to the diet's chemical composition and feed intake. Six Gobra zebu cows per management system were individually monitored over 10 months, diet and faeces were sampled each season and their chemical composition and dry matter intake (DMI) were predicted by near infrared spectroscopy. Each diet was fermented in vitro to assess methane production and volatile fatty acid concentration. The DMI and digestible organic matter intake (DOMI) decreased (P < 0.0001) during the dry seasons for both systems in the same range, but INT improved the crude protein of the diets (P < 0.0001). Enteric methane production (mmol.g-1 dry matter) was lower for TRAD than INT, except during the rainy season when TRAD cows experienced a higher increase (P = 0.002). The methanogenic potential (methane production in vitro × DMI) varied with the seasons and the system with more accentuated variations for TRAD (P < 0.0001). Methanogenic potential shows true reflection of the effects of the seasons and management systems. The results highlight that enteric methane emissions varied with seasonal changes and that intensifying the diet induced no mitigating effect.

Shifts in the rumen microbiota due to the type of carbohydrate and level of protein ingested by dairy cattle are associated with changes in rumen fermentation.

Balancing energy and nitrogen in the rumen is a key to both profitability and environmental sustainability. Four dairy cows were used in a Latin square experimental design to investigate the effect of severe nitrogen underfeeding (110 vs. 80% of requirements) and the type of carbohydrate consumed [neutral detergent fiber rich (FIB) vs. starch rich (STA)] on the rumen ecosystem. These dietary treatments modified both rumen fermentation and microbial populations. Compared with STA diets, consumption of FIB diets increased bacterial and fungal diversity in the rumen and also increased the concentrations of cellulolytic microorganisms, including protozoa (+38%), anaerobic fungi (+59%), and methanogens (+27%). This microbial adaptation to fiber utilization led to similar digestibility values for the 2 carbohydrate sources and was accompanied by a shift in the rumen fermentation patterns; when the FIB diets were consumed, the cows had greater ruminal pH, ammonia concentrations, and molar proportions of acetate and propionate compared with when they consumed the STA diets. Certain rumen microorganisms were sensitive to a shortage of nitrogen; rumen concentrations of ammonia were 49% lower when the low-protein (LP) diets were consumed as were total bacteria (-13%), anaerobic fungi (-28%), methanogens (-27%), protozoa (-19%), cellulolytic bacteria, and microbial diversity compared with when the high-protein (HP) diets were consumed. As a result, the digestibility of the LP diets was less than that of the HP diets. These findings demonstrated that the rumen microbial ecosystem is directly linked to the rumen fermentation pattern and, to some extent, to the efficiency of diet utilization by dairy cattle.

Effect of linseed fed as rolled seeds, extruded seeds or oil on fatty acid rumen metabolism and intestinal digestibility in cows.

Linseed, a source of linolenic acid, is used in ruminant diets to increase polyunsaturated fatty acids (FA) in animal products. Seed processing is known to have an impact on FA rumen metabolism, but few data are available for linseed. We studied the effect of linseed lipid on ruminal metabolism and intestinal digestibility in cows. Three modes of linseed processing: rolled linseed (RL), extruded linseed (EL) and linseed oil plus linseed meal (LO), supplemented at 7.5% of DM intake, were compared to a control diet (C). Duodenal flows, intestinal digestibility and plasma composition were determined. The duodenal flow of linolenic acid was similar among diets. The sum of t10 and t11-18:1, which were coeluted, was increased with lipid-supplemented diets and represented more than 60% of trans 18:1 for EL and LO diets. The main 18:2 isomers were c9, c12 and t11, c15 among the non-conjugated isomers, and t11, t13 among CLA. Linseed supplementation increased the duodenal flow of unsaturated intermediates of biohydrogenation, and this effect was more pronounced for extruded seeds and oil than for rolled seeds. For most 18-carbon FA, intestinal digestibility was slightly higher for C and LO diets than for RL and EL. Plasma concentrations of non-conjugated 18:2 and linolenic acid were similar among the lipid-supplemented diets. Within diet, profiles of 18:1 isomers (except c9) remained very similar between duodenal and plasma FA.

Methanogenic potential of tropical feeds rich in hydrolyzable tannins1,2.

The present study was carried out to determine the effect of Acacia nilotica, a tropical plant rich in hydrolyzable tannins (HT), on rumen fermentation and methane (CH4) production in vitro. We used leaves and pods from A. nilotica alone and combined. The combination of HT from A. nilotica leaves and pods and condensed tannins (CT) from Calliandra calothyrsus and Leucaena leucocephala were also evaluated to assess potential differences in biological activity between HT and CT. Four series of 24-h incubations were performed using rumen contents of 4 sheep fed a tropical grass (natural grassland based on Dichanthium spp.). A first experiment tested different levels of replacement of this tropical forage (control [CTL] without tannins) by A. nilotica leaves or pods: 0:100, 25:75, 50:50, 75:25 and 100:0. A second experiment tested the mixture of A. nilotica leaves and pods in different proportions: 100:0, 75:25, 50:50, 25:75, and 0:100. A third experiment tested the 50:50 combination of A. nilotica leaves or pods with C. calothyrsus and L. leucocephala. Acacia nilotica pods and leaves had a high content of HT (350 and 178 g/kg DM, respectively), whereas C. calothyrsus and L. leucocephala had a high content of CT (361 and 180 g/kg DM, respectively). The inclusion of HT from A. nilotica leaves and pods decreased CH4 production dose-dependently (P < 0.01). Total replacement of the CTL by A. nilotica decreased CH4 production by 64 and 55% with leaves and pods, respectively. Pods were richer in HT than leaves, but their antimethanogenic effect did not differ (P > 0.05). Although A. nilotica leaves and pods inhibited fermentation, as indicated by the lower gas production and VFA production (P < 0.01), this effect was less pronounced than for CH4. Volatile fatty acid production decreased by 12% in leaves and by 30% in pods when compared with the CTL alone. Positive associative effect was reported for VFA, when HT-rich sources and CT-rich sources were mixed. Combining the 2 sources of HT did not show associative effects on fermentation or CH4 production (P > 0.05). Hydrolyzable tannin-rich sources were more effective in suppressing methanogenesis than CT-rich sources. Our results show that HT-rich A. nilotica leaves and pods have the potential to reduce ruminal CH4 production.

Effectiveness of Interventions to Modulate the Rumen Microbiota Composition and Function in Pre-ruminant and Ruminant Lambs.

Modulating the assembly of the ruminal microbiota might have practical implications in production. We tested how an early-life dietary intervention in lambs influences the diversity and function of the ruminal microbiota during and after the intervention. Microbiota resilience during a repeated dietary intervention was also tested. The treatment, aiming to mitigate enteric methane emissions, combined garlic essential oil and linseed oil. Fifty-six lambs and their dams were allocated to two groups and treatment (T1) or placebo (C1) was drenched from birth until 10 weeks of life. Lambs were weaned at 8 weeks. From 16 to 20 weeks, lambs in each group were divided in two subgroups that received (T1-T2 and C1-T2) or not (T1-C2 and C1-C2) the same treatment. Measurements were done at 8, 14, and 20 weeks. Average daily gain was similar between groups. Methane production was reduced by treatment at 8 and 20 weeks but at 14 weeks it was similar between C1 and T1. Interestingly, early-life treated lambs displayed a numerical increase (P = 0.12) in methane emissions at 20 weeks compared with non-treated lambs. Concentration of VFA was not affected by the intervention at 8 or 14 weeks but a lower concentration was observed in T2 lambs compared with C2 at week 20. Metataxonomics (rRNA gene) revealed differences in archaeal communities between groups of lambs when treatment was applied (weeks 8 and 20); whereas, in accord with methane emissions, these differences disappeared when treatment was discontinued (week 14). Protozoal community structure was not affected by treatment. In contrast, bacterial community structure differed between treated and non-treated lambs during and after the intervention. Rumen and urine LC-MS and NMR metabolomics at week 20 separated C2 from T2 lambs and correlation analysis highlighted interactions between microbes and metabolites, notably that of methylated compounds and Methanomassiliicocceae methanogens. This study demonstrates that a long-term early-life intervention induced modifications in the composition of the rumen bacterial community that persisted after the intervention ceased with little or no effect on archaeal and protozoal communities. However, there was no persistency of the early-life intervention on methanogenesis indicating resilience for this function.

Dynamic features of the rumen metabolism of linoleic acid, linolenic acid and linseed oil measured in vitro.

The lipid quality of ruminant products is largely determined by the extent of rumen microbial biohydrogenation (BH) of polyunsaturated fatty acids (FAs) and the substances formed thereby. In vitro batch incubations with mixed rumen bacteria were tracked over 24 h to characterize the profiles and kinetics of the BH products from three lipid sources: pure linoleic acid (c9,c12-18:2), pure linolenic acid (c9,c12,c15-18:3) and linseed oil (mainly c9,c12,c15-18:3 in triacylglycerols). After 24 h of incubation biohydrogenation was more complete for c9,c12-18:2, which gave mainly stearic acid (18:0), than for c9,c12,c15-18:3, which yielded mainly trans-18:1 FAs. This suggests inhibition of the final BH step (18:1 to 18:0). Incubations of c9,c12-18:2 resulted in high levels of carbon 10- and 12-desaturated 18:1, t10,c12- and c9,t11-CLAs. Incubations of c9,c12,c15-18:3 resulted in high levels of t11-18:1, carbon 13- and 15-desaturated 18:1 as well as t11,c15-18:2 and 11,13-CLAs. A comparative study of linolenic acid and linseed oil kinetics revealed that the BH process was not significantly slowed by the esterification of polyunsaturated FAs, but may have been limited by the isomerization step in which the cis12 double bond goes to the trans11 position. The disappearance rates of c9,c12-18:2 and c9,c12,c15-18:3 ranged from 23.6 to 44.6%/h. The wide variety of BH intermediates found here underlines the large number of possible BH pathways. These data help provide a basis for dynamic approaches to BH processes.

The Structural and Functional Capacity of Ruminal and Cecal Microbiota in Growing Cattle Was Unaffected by Dietary Supplementation of Linseed Oil and Nitrate.

Microorganisms in the digestive tract of ruminants differ in their functionality and ability to use feed constituents. While cecal microbiota play an important role in post-rumen fermentation of residual substrates undigested in the rumen, limited knowledge exists regarding its structure and function. In this trial we investigated the effect of dietary supplementation with linseed oil and nitrate on methane emissions and on the structure of ruminal and cecal microbiota of growing bulls. Animals were allocated to either a CTL (control) or LINNIT (CTL supplemented with 1.9% linseed and 1.0% nitrates) diet. Methane emissions were measured using the GreenFeed system. Microbial diversity was assessed using amplicon sequencing of microbial genomic DNA. Additionally, total RNA was extracted from ruminal contents and functional mcrA and mtt genes were targeted in amplicon sequencing approach to explore the diversity of functional gene expression in methanogens. LINNIT had no effect on methane yield (g/kg DMI) even though it decreased methane production by 9% (g/day; P < 0.05). Methanobrevibacter- and Methanomassiliicoccaceae-related OTUs were more abundant in cecum (72 and 24%) compared to rumen (60 and 11%) irrespective of the diet (P < 0.05). Feeding LINNIT reduced the relative abundance of Methanomassiliicoccaceae mcrA cDNA reads in the rumen. Principal component analysis revealed significant differences in taxonomic composition and abundance of bacterial communities between rumen and cecum. Treatment decreased the relative abundance of a few Ruminococcaceae genera, without affecting global bacterial community structure. Our research confirms a high level of heterogeneity in species composition of microbial consortia in the main gastrointestinal compartments where feed is fermented in ruminants. There was a parallel between the lack of effect of LINNIT on ruminal and cecal microbial community structure and functions on one side and methane emission changes on the other. These results suggest that the sequencing strategy used here to study microbial diversity and function accurately reflected the absence of effect on methane phenotypes in bulls treated with linseed plus nitrate.

An assessment of the impact of rumenocentesis on pain and stress in cattle and the effect of local anaesthesia.

Rumenocentesis is commonly used to collect rumen fluid to screen for acidosis. This study was designed to investigate whether rumenocentesis induced pain and stress and, if so, whether local anaesthesia could limit this. Twenty-four dairy cows were assigned to one of three treatments: (1) rumenocentesis with local anaesthesia (AR); (2) rumenocentesis without local anaesthesia (R); and (3) local anaesthesia only (A). Treatments were performed in a restraining cage. The cows were placed in the cage on three consecutive days and anaesthesia and/or rumenocentesis was performed on the second day. Blood samples for cortisol determination and heart rate were recorded from 0.25 h before treatment until 4h after. Behaviour was noted while the cows were caged. Feed intake and milk production were measured the week before treatment, on the day of treatment, and the day after. With all three treatments, cortisol concentrations and heart rate were increased while cows were in the cage. Cortisol, cardiac and behavioural responses were not significantly higher in the R and AR treatments than the A group. Cortisol concentrations and heart rate did not change between days. Feed intake and milk production were unaffected by the treatments. It was concluded that rumenocentesis does not appear more stressful than local anaesthesia or handling.

t10,c12-18:2-induced milk fat depression is less pronounced in cows fed high-concentrate diets.

In intensively reared dairy cows, milk fat secretion is reduced in response to high-concentrate diets and it is often referred to as the "milk fat depression" (MFD) syndrome. Some trans fatty acid (FA) isomers produced in the rumen of the cows, including t10,c12-18:2, are known for their inhibitory effect on mammary lipogenesis. To study whether this effect depends on the basal diet, duodenal infusions of t10,c12-18:2 were performed on cows fed four different diets (a factorial arrangement of forage:concentrate ratio and linseed oil supplementation). The overall response obtained with t10,c12-18:2 infusion was consistent with previous studies: a decrease in milk fat content and yield without significant variations in milk yield. Mean transfer efficiency of infused t10,c12-18:2 was 19.6%. However, the decrease in milk fat and FA yields (both de novo synthesis and preformed long-chain FA) was less pronounced in cows fed high-concentrate diets (-27% of the initial level), compared with cows fed low-concentrate diets (-42% of initial level). This difference was independent of dietary oil supplementation and milk FA yield before infusion. Results pertaining to effects of dietary forage:concentrate ratio were confirmed by statistical meta-analysis of data from previously published t10,c12-18:2 infusion experiments. This study shows that in cows fed MFD diets the mammary gland becomes more resistant to or experiences a lower response potential to further inhibition of lipogenesis and/or delta-9 desaturation of FA.

Variation in content and composition of phenolic compounds in permanent pastures according to botanical variation.

Phenolic compounds contribute to the micronutrient composition of pasture, which in turn may affect animal product composition. To assess the importance and variations in content of these compounds, the polyphenolic and botanical compositions of 24 permanent pastures located in one lowland and two upland regions were studied at equivalent stages of growth. Phenolic fractions were characterized and quantified using HPLC-PDA-ESI-QToF, and the total content was determined by colorimetry over each whole pasture. A rise in altitude was accompanied by a marked increase in total phenolic content, linked to changes in botanical composition, but did not have any influence on the distribution according to molecular class. For all of the pastures, significantly different patterns due to qualitative and quantitative differences among the 92 separate peaks were observed with 31 compounds identified. The involvement of certain plants in the variations of content and composition in phenolic compounds of pastures was statistically evaluated.